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Detection of Y‐bearing spermatozoa by DNA‐DNA in situ hybridisation
Author(s) -
West John D.,
West Katrine M.,
Aitken R. John
Publication year - 1989
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.1080010308
Subject(s) - biology , dna , in situ hybridisation , sperm , chromatin , y chromosome , microbiology and biotechnology , chromosome , hybridization probe , genetics , gene
In situ hybridisation of a Y chromosome‐specific DNA probe to preparations of decondensed spermatozoa revealed approximately 46.7% labelled spermatozoa among 3,900 scored. This is not significantly different from the 50% expected if only the Y chromosome‐bearing spermatozoa are hybridised. Control hybridisations of Escherichia coli DNA and salmon testis DNA to decondensed sperm produced no significant labelling, whereas more than 99% of the spermatozoa were heavily labelled after hybridisation to total human DNA. These controls indicate that the methodology described in this paper renders the chromatin accessible for hybridisation and that the 50% hybridisation observed with the Y chromosome DNA probe was specific. In situ hybridisation with the Y probe therefore identifies the Y‐bearing spermatozoa, and the protocol described should prove useful in evaluating methods of separating Y‐bearing and X‐bearing spermatozoa.